s-Triazine compounds, pharmaceutical compositions and method of using the same

ABSTRACT

Novel s-triazine compounds are disclosed that have a formula represented by the following: 
                         
The compounds may be prepared as pharmaceutical compositions, and may be used for the prevention and treatment of a variety of conditions in mammals including humans, where microbial infection is either a direct cause or a related condition.

RELATED APPLICATION

The present application claims the benefit under 35 U.S.C. §119 of U.S.Provisional Application Ser. No. 60/931,514 filed May 23, 2007, thecontents of which application is hereby incorporated by reference in itsentirety.

GOVERNMENT SUPPORT

This invention was made at least in part, with government support underGrant No. N00014-03-1-0129 awarded by the Office of Naval Research.Accordingly, the United States Government has certain rights in theinvention.

FIELD OF THE INVENTION

This invention relates to novel compositions containing actives-triazine compounds, and particularly, such s-triazine compounds asdemonstrate antimicrobial, antifungal or antiviral activity. Theinvention also relates to methods for the preparation of the peptidecompositions, and their use in preventing and/or treating conditionsresulting from the unwanted presence of microbial, fungal or viralactivity.

BACKGROUND OF THE INVENTION

As multidrug-resistant bacterial strains emerge in increasing numbers,the need for new kinds of antibiotics is growing. For the last fewdecades it has been found that a wide range of antimicrobial peptidesare secreted by multicellular organisms in response to infection byforeign bacteria, viruses, or fungi (1-4). These form part of the innateimmune response to infection, which is short term and fast actingrelative to humoral immunity (3). These peptides have been considered asprospective antibiotic agents because their effect is rapid, broadspectrum and indifferent to resistance to standard antibiotics such aspenicillin (5-6). Antimicrobial peptides differ dramatically in size,sequence and structure, apparently sharing only amphipathic characterand positive charge (1, 5). The proposed mechanisms of action ofantimicrobial peptides commonly focus on the interaction between thesepeptides and the plasma membrane of bacterial cells, even though manyantimicrobial peptides also employ more sophisticated mechanisms (7).Recently, the pharmocophore of short cationic antimicrobial peptides hasbeen extensively studied and the results showed that short cationicpeptides consisting of only Arginine (R) and Tryptophan (W) could serveas moderately effective antimicrobial agents (8-9).

Besides various advantages over conventional antibiotics, the practicaluse of antimicrobial peptides, however, are limited by many factors.These peptides are usually more expensive to make, vulnerable toprotease degradation, and have relatively high toxicity. A number ofnonnatural peptides built from beta-amino acids or peptoids, as well asother peptide mimics have been studied in order to overcome theseproblems (10-12). In this study, we designed and screened inexpensivesmall compounds to mimic the hydrophobic-cationic pattern observed inthe pharmocophore of small cationic antimicrobial peptides using1,3,5-triazine as a template. Previous studies showed that possibleantimicrobials could be identified through combinatorial librariesconstructed to have varieties of tri-substituted 1,3,5-triazines (13).

From the above, it remains that a continuing need exists for thestepwise design and optimization of different functional groups (mainlyhydrophobic, bulky or charged groups) on the triazine scaffold in searchof potential new antimicrobials, as well as to gain insight as to thestructure-function relationship of these agents.

Furthermore, it remains that a continuing need exists for thedevelopment of modalities that can deliver effective antibiotics-triazine compounds in a manner that confers both improved stabilityand economy of the therapeutic, but importantly, significantly improvesthe therapeutic efficacy and strength of the resultant molecule. It istoward the fulfillment of these and other related objectives that thepresent invention is directed.

SUMMARY OF THE INVENTION

As set forth earlier herein, the present invention relates to s-triazinecompounds, pharmaceutical compositions thereof and their use asantimicrobial/antiviral/antifungal compounds.

The present invention relates to s-triazine compounds havingantimicrobial properties, according to formula I:

wherein

each R¹, R^(1a) and R² is independently substituted amino; or apharmaceutically acceptable salt, solvate or prodrug thereof; andstereoisomers, isotopic variants and tautomers thereof.

More particularly, the present invention relates to s-triazine compoundshaving antimicrobial properties, according to formula Ia:

wherein

-   -   each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f), R^(2g),        R^(2h) and R^(2j) is independently selected from hydrogen, and        substituted or unsubstituted C—C₆ alkyl; or any two of R^(2c),        R^(2d), R^(2e), R^(2f) and R^(2g) may join together to form a        cycloalkyl or cycloheteroalkyl ring or 3-7 atoms;    -   R³ is R⁴ or substituted or unsubstituted C₁-C₆ alkyl; or is        selected from a 3-13 membered cycloalkyl, heterocycloalkyl, aryl        and heteroaryl ring system, which can be optionally substituted        with one or more substituents independently selected from halo,        hydroxyl, amino, cyano, sulfo, sulfanyl, sulfinyl, amido,        carboxy, ester, alkyl, substituted alkyl, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, and sulfonamido;    -   R⁴ is

-   -   wherein C¹, C² and C³ taken together with the C atom to which        they are attached to form a bi- or tri-cyloalkyl or        cycloheteroalkyl ring system of 7-13 atoms; and wherein the ring        system is substituted or unsubstituted;    -   m is 0, 1, 2 or 3; and n is 2, 3 or 4;    -   or a pharmaceutically acceptable salt, solvate or prodrug        thereof;    -   and stereoisomers, isotopic variants and tautomers thereof.

In one embodiment, with respect to compounds of formula Ia, R⁴ issubstituted or unsubstituted

In a preferred embodiment, with respect to compounds of formula Ia, R⁴is adamantane.

In a preferred embodiment, with respect to s-triazine compounds offormula Ia, each R^(2b), R^(2d), R^(2e), R^(2h) and R^(2j) is hydrogen.

In one embodiment, with respect to s-triazine compounds of formula Ia,each R^(2f) and R^(2g) is Me, Et or n-Bu. In a preferred embodiment,each R^(2f) and R^(2g) is n-Bu.

In one embodiment, with respect to s-triazine compounds of formula Ia, nis 2 or 3. In a preferred embodiment, n is 3.

In one embodiment, with respect to s-triazine compounds of formula Ia, mis 0, 1 or 2.

In one embodiment, with respect to s-triazine compounds of formula Ia,R⁴ is cycloheptyl, cyclohexyl or phenyl.

In a further aspect, the present invention provides the method for thepreparation of the s-triazine compounds of the invention.

In a further aspect, the s-triazine compounds of the invention may beused to treat microbial or fungal conditions affecting lower animals,and possibly, plants. The s-triazine compounds could be designed andassembled to include the s-triazine compounds pertinent for thetreatment of a particular microbe or fungus of interest, and thenformulated into appropriate compositions and dosage forms foradministration or application to an affected host.

In a further aspect, the present invention provides pharmaceuticalcompositions comprising a peptide of the invention, and a pharmaceuticalcarrier, excipient or diluent. In this aspect of the invention, thepharmaceutical composition can comprise one or more variant s-triazinecompounds of the invention, prepared, for example, with a differingarray of peptide linkers, to afford a more comprehensive treatment inthe instance where a multiplicity of microbial/viral/fungal antigens areknown to be present.

In a further aspect of the invention, a method is disclosed for treatingmammals, including humans, as well as lower mammalian species,susceptible to or afflicted with a condition attributable to orresulting from amicrobial, viral or fungal infection, which methodcomprises administering an effective amount of a pharmaceuticalcomposition containing or comprising the s-triazine compounds justdescribed.

In additional aspects, this invention provides methods for synthesizingthe compounds of the invention, with representative synthetic protocolsand pathways disclosed later on herein.

Other objects and advantages will become apparent to those skilled inthe art from a consideration of the ensuing detailed description, whichproceeds with reference to the following illustrative drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 graphically depicts the results of the measurement of the abilityof a triazine compound of the invention to induce leakage of calcein,entrapped within the interior of large unilamellar vesicles of modelmembrane POPG. The extent of leakage of encapsulated calcein wasdetected by its fluorescence at 515 nm, with 100% leakage calibrated byaddition of 0.2% Triton X-100.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

When describing the compounds, pharmaceutical compositions containingsuch compounds and methods of using such compounds and compositions, thefollowing terms have the following meanings unless otherwise indicated.It should also be understood that any of the moieties defined forthbelow may be substituted with a variety of substituents, and that therespective definitions are intended to include such substituted moietieswithin their scope. It should be further understood that the terms“groups” and “radicals” can be considered interchangeable when usedherein.

“Acyl” refers to a radical —C(O)R²⁰, where R²⁰ is hydrogen, alkyl,cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl,heteroarylalkyl as defined herein. Representative examples include, butare not limited to, formyl, acetyl, cylcohexylcarbonyl,cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl and the like.

“Acylamino” refers to a radical —NR²¹C(O)R²², where R²¹ is hydrogen,alkyl, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl,heteroaryl, heteroarylalkyl and R²² is hydrogen, alkyl, alkoxy,cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroarylor heteroarylalkyl, as defined herein. Representative examples include,but are not limited to, formylamino, acetylamino,cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino,benzylcarbonylamino and the like.

“Acyloxy” refers to the group —OC(O)R²³ where R²³ is hydrogen, alkyl,aryl or cycloalkyl.

“Substituted alkenyl” includes those groups recited in the definition of“substituted” herein, and particularly refers to an alkenyl group having1 or more substituents, for instance from 1 to 5 substituents, andparticularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Alkoxy” refers to the group —OR²⁴ where R²⁴ is alkyl. Particular alkoxygroups include, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy,1,2-dimethylbutoxy, and the like.

“Substituted alkoxy” includes those groups recited in the definition of“substituted” herein, and particularly refers to an alkoxy group having1 or more substituents, for instance from 1 to 5 substituents, andparticularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,heteroaryl, hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy,thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— andaryl-S(O)₂—.

“Alkoxycarbonylamino” refers to the group —NR²⁵C(O)OR²⁶, where R²⁵ ishydrogen, alkyl, aryl or cycloalkyl, and R²⁶ is alkyl or cycloalkyl.

“Alkyl” refers to monovalent saturated alkane radical groupsparticularly having up to about 11 carbon atoms, more particularly as alower alkyl, from 1 to 8 carbon atoms and still more particularly, from1 to 6 carbon atoms. The hydrocarbon chain may be eitherstraight-chained or branched. This term is exemplified by groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, tert-butyl,n-hexyl, n-octyl, tert-octyl and the like. The term “lower alkyl” refersto alkyl groups having 1 to 6 carbon atoms. The term “alkyl” alsoincludes “cycloalkyls” as defined below.

“Substituted alkyl” includes those groups recited in the definition of“substituted” herein, and particularly refers to an alkyl group having 1or more substituents, for instance from 1 to 5 substituents, andparticularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, heteroaryl, keto, nitro, thioalkoxy, substituted thioalkoxy,thioaryloxy, thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂—, andaryl-S(O)₂—.

“Alkylene” refers to divalent saturated alkene radical groups having 1to 11 carbon atoms and more particularly 1 to 6 carbon atoms which canbe straight-chained or branched. This term is exemplified by groups suchas methylene (—CH₂—), ethylene (—CH₂CH₂—), the propylene isomers (e.g.,—CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like.

“Substituted alkylene” includes those groups recited in the definitionof “substituted” herein, and particularly refers to an alkylene grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, amino-carbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, halogen, hydroxyl, keto, nitro, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioketo, thiol, alkyl-S(O)—,aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Alkenyl” refers to monovalent olefinically unsaturated hydrocarbylgroups preferably having 2 to 11 carbon atoms, particularly, from 2 to 8carbon atoms, and more particularly, from 2 to 6 carbon atoms, which canbe straight-chained or branched and having at least 1 and particularlyfrom 1 to 2 sites of olefinic unsaturation. Particular alkenyl groupsinclude ethenyl (—CH═CH₂), n-propenyl (—CH₂CH═CH₂), isopropenyl(—C(CH₃)═CH₂), vinyl and substituted vinyl, and the like.

“Alkenylene” refers to divalent olefinically unsaturated hydrocarbylgroups particularly having up to about 11 carbon atoms and moreparticularly 2 to 6 carbon atoms which can be straight-chained orbranched and having at least 1 and particularly from 1 to 2 sites ofolefinic unsaturation. This term is exemplified by groups such asethenylene (—CH═CH—), the propenylene isomers (e.g., —CH═CHCH₂— and—C(CH₃)═CH— and —CH═C(CH₃)—) and the like.

“Alkynyl” refers to acetylenically or alkynically unsaturatedhydrocarbyl groups particularly having 2 to 11 carbon atoms, and moreparticularly 2 to 6 carbon atoms which can be straight-chained orbranched and having at least 1 and particularly from 1 to 2 sites ofalkynyl unsaturation. Particular non-limiting examples of alkynyl groupsinclude acetylenic, ethynyl (—C≡CH), propargyl (—CH₂C≡CH), and the like.

“Substituted alkynyl” includes those groups recited in the definition of“substituted” herein, and particularly refers to an alkynyl group having1 or more substituents, for instance from 1 to 5 substituents, andparticularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Alkanoyl” or “acyl” as used herein refers to the group R²⁷—C(O)—, whereR²⁷ is hydrogen or alkyl as defined above.

“Aryl” refers to a monovalent aromatic hydrocarbon group derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. Typical aryl groups include, but are not limitedto, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexylene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene and the like. Particularly, anaryl group comprises from 6 to 14 carbon atoms.

“Substituted Aryl” includes those groups recited in the definition of“substituted” herein, and particularly refers to an aryl group that mayoptionally be substituted with 1 or more substituents, for instance from1 to 5 substituents, particularly 1 to 3 substituents, selected from thegroup consisting of acyl, acylamino, acyloxy, alkenyl, substitutedalkenyl, alkoxy, substituted alkoxy, alkoxycarbonyl, alkyl, substitutedalkyl, alkynyl, substituted alkynyl, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thiol,alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Fused Aryl” refers to an aryl having two of its ring carbon in commonwith a second aryl ring or with an aliphatic ring.

“Alkaryl” refers to an aryl group, as defined above, substituted withone or more alkyl groups, as defined above.

“Aralkyl” or “arylalkyl” refers to an alkyl group, as defined above,substituted with one or more aryl groups, as defined above.

“Aryloxy” refers to —O-aryl groups wherein “aryl” is as defined above.

“Alkylamino” refers to the group alkyl-NR²⁸R²⁹, wherein each of R²⁸ andR²⁹ are independently selected from hydrogen and alkyl.

“Arylamino” refers to the group aryl-NR³⁰R³¹, wherein each of R³⁰ andR³¹ are independently selected from hydrogen, aryl and heteroaryl.

“Alkoxyamino” refers to a radical —N(H)OR³² where R³² represents analkyl or cycloalkyl group as defined herein.

“Alkoxycarbonyl” refers to a radical —C(O)-alkoxy where alkoxy is asdefined herein.

“Alkylarylamino” refers to a radical —NR³³R³⁴ where R³³ represents analkyl or cycloalkyl group and R³⁴ is an aryl as defined herein.

“Alkylsulfonyl” refers to a radical —S(O)₂R³⁵ where R³⁵ is an alkyl orcycloalkyl group as defined herein. Representative examples include, butare not limited to, methylsulfonyl, ethylsulfonyl, propylsulfonyl,butylsulfonyl and the like.

“Alkylsulfinyl” refers to a radical —S(O)R³⁵ where R³⁵ is an alkyl orcycloalkyl group as defined herein. Representative examples include, butare not limited to, methylsulfinyl, ethylsulfinyl, propylsulfinyl,butylsulfinyl and the like.

“Alkylthio” refers to a radical —SR³⁵ where R³⁵ is an alkyl orcycloalkyl group as defined herein that may be optionally substituted asdefined herein. Representative examples include, but are not limited to,methylthio, ethylthio, propylthio, butylthio, and the like.

“Amino” refers to the radical —NH₂.

“Substituted amino” includes those groups recited in the definition of“substituted” herein, and particularly refers to the group —N(R³⁶)₂where each R³⁶ is independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, aryl, cycloalkyl, substituted cycloalkyl,and where both R groups are joined to form an alkylene group. When bothR groups are hydrogen, —N(R³⁶)₂ is an amino group.

“Aminocarbonyl” refers to the group —C(O)NR³⁷R³⁷ where each R³⁷ isindependently hydrogen, alkyl, aryl and cycloalkyl, or where the R³⁷groups are joined to form an alkylene group.

“Aminocarbonylamino” refers to the group —NR³⁸C(O)NR³⁸R³⁸ where each R³⁸is independently hydrogen, alkyl, aryl or cycloalkyl, or where two Rgroups are joined to form an alkylene group.

“Aminocarbonyloxy” refers to the group —OC(O)NR³⁹R³⁹ where each R³⁹ isindependently hydrogen, alkyl, aryl or cycloalkyl, or where the R groupsare joined to form an alkylene group.

“Arylalkyloxy” refers to an —O-arylalkyl radical where arylalkyl is asdefined herein.

“Arylamino” means a radical —NHR⁴⁰ where R⁴⁰ represents an aryl group asdefined herein.

“Aryloxycarbonyl” refers to a radical —C(O)—O-aryl where aryl is asdefined herein.

“Arylsulfonyl” refers to a radical —S(O)₂R⁴¹ where R⁴¹ is an aryl orheteroaryl group as defined herein.

“Azido” refers to the radical —N₃.

“Bicycloaryl” refers to a monovalent aromatic hydrocarbon group derivedby the removal of one hydrogen atom from a single carbon atom of aparent bicycloaromatic ring system. Typical bicycloaryl groups include,but are not limited to, groups derived from indane, indene, naphthalene,tetrahydronaphthalene, and the like. Particularly, an aryl groupcomprises from 8 to 11 carbon atoms.

“Bicycloheteroaryl” refers to a monovalent bicycloheteroaromatic groupderived by the removal of one hydrogen atom from a single atom of aparent bicycloheteroaromatic ring system. Typical bicycloheteroarylgroups include, but are not limited to, groups derived from benzofuran,benzimidazole, benzindazole, benzdioxane, chromene, chromane, cinnoline,phthalazine, indole, indoline, indolizine, isobenzofuran, isochromene,isoindole, isoindoline, isoquinoline, benzothiazole, benzoxazole,naphthyridine, benzoxadiazole, pteridine, purine, benzopyran,benzpyrazine, pyridopyrimidine, quinazoline, quinoline, quinolizine,quinoxaline, benzomorphan, tetrahydroisoquinoline, tetrahydroquinoline,and the like. Preferably, the bicycloheteroaryl group is between 9-11membered bicycloheteroaryl, with 5-10 membered heteroaryl beingparticularly preferred. Particular bicycloheteroaryl groups are thosederived from benzothiophene, benzofuran, benzothiazole, indole,quinoline, isoquinoline, benzimidazole, benzoxazole and benzdioxane.

“Carbamoyl” refers to the radical —C(O)N(R⁴²)₂ where each R⁴² group isindependently hydrogen, alkyl, cycloalkyl or aryl, as defined herein,which may be optionally substituted as defined herein.

“Carboxy” refers to the radical —C(O)OH.

“Carboxyamino” refers to the radical —N(H)C(O)OH.

“Cycloalkyl” refers to cyclic hydrocarbyl groups having from 3 to about10 carbon atoms and having a single cyclic ring or multiple condensedrings, including fused and bridged ring systems, which optionally can besubstituted with from 1 to 3 alkyl groups. Such cycloalkyl groupsinclude, by way of example, single ring structures such as cyclopropyl,cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl,2-methylcyclopentyl, 2-methylcyclooctyl, and the like, and multiple ringstructures such as adamantanyl, and the like.

“Substituted cycloalkyl” includes those groups recited in the definitionof “substituted” herein, and particularly refers to a cycloalkyl grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Cycloalkoxy” refers to the group —OR⁴³ where R⁴³ is cycloalkyl. Suchcycloalkoxy groups include, by way of example, cyclopentoxy, cyclohexoxyand the like.

“Cycloalkenyl” refers to cyclic hydrocarbyl groups having from 3 to 10carbon atoms and having a single cyclic ring or multiple condensedrings, including fused and bridged ring systems and having at least oneand particularly from 1 to 2 sites of olefinic unsaturation. Suchcycloalkenyl groups include, by way of example, single ring structuressuch as cyclohexenyl, cyclopentenyl, cyclopropenyl, and the like.

“Substituted cycloalkenyl” includes those groups recited in thedefinition of “substituted” herein, and particularly refers to acycloalkenyl group having 1 or more substituents, for instance from 1 to5 substituents, and particularly from 1 to 3 substituents, selected fromthe group consisting of acyl, acylamino, acyloxy, alkoxy, substitutedalkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Fused Cycloalkenyl” refers to a cycloalkenyl having two of its ringcarbon atoms in common with a second aliphatic or aromatic ring andhaving its olefinic unsaturation located to impart aromaticity to thecycloalkenyl ring.

“Cyanato” refers to the radical —OCN.

“Cyano” refers to the radical —CN.

“Dialkylamino” means a radical —NR⁴⁴R⁴⁵ where R⁴⁴ and R⁴⁵ independentlyrepresent an alkyl, substituted alkyl, aryl, substituted aryl,cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substitutedcycloheteroalkyl, heteroaryl, or substituted heteroaryl group as definedherein.

“Ethenyl” refers to substituted or unsubstituted —(C═C)—.

“Ethylene” refers to substituted or unsubstituted —(C—C)—.

“Ethynyl” refers to —(C≡C)—.

“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo. Preferredhalo groups are either fluoro or chloro.

“Hydroxy” refers to the radical —OH.

“Nitro” refers to the radical —NO₂.

“Substituted” refers to a group in which one or more hydrogen atoms areeach independently replaced with the same or different substituent(s).Typical substituents include, but are not limited to, —X, —R⁴⁶, —O⁻, ═O,—OR⁴⁶, —SR⁴⁶, —S⁻, ═S⁻, —NR⁴⁶R⁴⁷, ═NR⁴⁶, —CX₃, —CF₃, —CN, —OCN, —SCN,—NO, —NO₂, ═N₂, —N₃, —S(O)₂O—, —S(O)₂OH, —S(O)₂R⁴⁶, —OS(O₂)O⁻,—OS(O)₂R⁴⁶, —P(O)(O⁻)₂, —P(O)(OR⁴⁶)(O⁻), —OP(O)(OR⁴⁶)(OR⁴⁷), —C(O)R⁴⁶,—C(S)R⁴⁶, —C(O)OR⁴³, —C(O)NR⁴⁶R⁴⁷, —C(O)O⁻, —C(S)OR⁴⁶, —NR⁴⁸C(O)NR⁴⁶R⁴⁷,—NR⁴⁸C(S)NR⁴⁶R⁴⁷, —NR⁴⁹C(NR⁴⁸)NR⁴⁶R⁴⁷ and —C(NR⁴⁸)NR⁴⁶R⁴⁷ where each Xis independently a halogen; each R⁴⁶, R⁴⁷, R⁴⁸ and R⁴⁹ are independentlyhydrogen, alkyl, substituted alkyl, aryl, substituted alkyl, arylalkyl,substituted alkyl, cycloalkyl, substituted alkyl, cycloheteroalkyl,substituted cycloheteroalkyl, heteroalkyl, substituted heteroalkyl,heteroaryl, substituted heteroaryl, heteroarylalkyl, substitutedheteroarylalkyl, —NR⁵⁰R⁵¹, —C(O)R⁵⁰ or —S(O)₂R⁵⁰ or optionally R⁵⁰ andR⁵¹ together with the atom to which they are both attached form acycloheteroalkyl or substituted cycloheteroalkyl ring; and R⁵⁰ and R⁵¹are independently hydrogen, alkyl, substituted alkyl, aryl, substitutedalkyl, arylalkyl, substituted alkyl, cycloalkyl, substituted alkyl,cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substitutedheteroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl orsubstituted heteroarylalkyl.

Examples of representative substituted aryls include the following:

In these formulae one of R⁵² and R⁵³ may be hydrogen and at least one ofR⁵² and R⁵³ is each independently selected from alkyl, alkenyl, alkynyl,cycloheteroalkyl, alkanoyl, alkoxy, aryloxy, heteroaryloxy, alkylamino,arylamino, heteroarylamino, NR⁵⁴COR⁵⁵, NR⁵⁴SOR⁵⁵, NR⁵⁴SO₂R⁵⁷, COOalkyl,COOaryl, CONR⁵⁴R⁵⁵, CONR⁵⁴OR⁵⁵, NR⁵⁴R⁵⁵, SO₂NR⁵⁴R⁵⁵, S-alkyl, S-alkyl,SOalkyl, SO₂alkyl, Saryl, SOaryl, SO₂aryl; or R⁵² and R⁵³ may be joinedto form a cyclic ring (saturated or unsaturated) from 5 to 8 atoms,optionally containing one or more heteroatoms selected from the group N,O or S. R⁵⁴, R⁵⁵, and R⁵⁶ are independently hydrogen, alkyl, alkenyl,alkynyl, perfluoroalkyl, cycloalkyl, cycloheteroalkyl, aryl, substitutedaryl, heteroaryl, substituted or hetero alkyl or the like.

“Hetero” when used to describe a compound or a group present on acompound means that one or more carbon atoms in the compound or grouphave been replaced by a nitrogen, oxygen, or sulfur heteroatom. Heteromay be applied to any of the hydrocarbyl groups described above such asalkyl, e.g. heteroalkyl, cycloalkyl, e.g. cycloheteroalkyl, aryl, e.g.heteroaryl, cycloalkenyl, cycloheteroalkenyl, and the like having from 1to 5, and especially from 1 to 3 heteroatoms.

“Heteroaryl” refers to a monovalent heteroaromatic group derived by theremoval of one hydrogen atom from a single atom of a parentheteroaromatic ring system. Typical heteroaryl groups include, but arenot limited to, groups derived from acridine, arsindole, carbazole,β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole,indole, indoline, indolizine, isobenzofuran, isochromene, isoindole,isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine,oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline,phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, quinazoline,quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,thiophene, triazole, xanthene, and the like. Preferably, the heteroarylgroup is between 5-15 membered heteroaryl, with 5-10 membered heteroarylbeing particularly preferred. Particular heteroaryl groups are thosederived from thiophene, pyrrole, benzothiophene, benzofuran, indole,pyridine, quinoline, imidazole, oxazole and pyrazine.

Examples of representative heteroaryls include the following:

wherein each Y is selected from carbonyl, N, NR⁵⁸, O, and S; and R⁵⁸ isindependently hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl,heteroaryl, heteroalkyl or the like.

As used herein, the term “cycloheteroalkyl” refers to a stableheterocyclic non-aromatic ring and fused rings containing one or moreheteroatoms independently selected from N, O and S. A fused heterocyclicring system may include carbocyclic rings and need only include oneheterocyclic ring. Examples of heterocyclic rings include, but are notlimited to, piperazinyl, homopiperazinyl, piperidinyl and morpholinyl,and are shown in the following illustrative examples:

wherein each X is selected from CR⁵⁸, CR⁵⁸ ₂, NR⁵⁸, O and S; and each Yis selected from NR⁵⁸, O and S; and R⁵⁸ is independently hydrogen,alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, heteroalkyl orthe like. These cycloheteroalkyl rings may be optionally substitutedwith one or more groups selected from the group consisting of acyl,acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl,alkoxycarbonylamino, amino, substituted amino, aminocarbonyl,aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl,cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxyl, keto,nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, thiol,alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—. Substitutinggroups include carbonyl or thiocarbonyl which provide, for example,lactam and urea derivatives.

Examples of representative cycloheteroalkenyls include the following:

wherein each X is selected from CR⁵⁸, CR⁵⁸ ₂, NR⁵⁸, O and S; and each Yis selected from carbonyl, N, NR⁵⁸, O and S; and R⁵⁸ is independentlyhydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,heteroalkyl or the like.

Examples of representative aryl having hetero atoms containingsubstitution include the following:

wherein each X is selected from C—R⁵⁸ ₂, NR⁵⁸, O and S; and each Y isselected from carbonyl, NR⁵⁸, O and S; and R⁵⁸ is independentlyhydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl,heteroalkyl or the like.

“Hetero substituent” refers to a halo, O, S or N atom-containingfunctionality that may be present as an R group present as substituentsdirectly on the ring atoms of the compounds provided herein or may bepresent as a substituent in the “substituted” aryl and aliphatic groupspresent in the compounds.

Examples of hetero substituents include:

-halo,

—NO₂, —NH₂, —NHR⁵⁹, —N(R⁵⁹)₂,

—NRCOR, —NR⁵⁹SOR⁵⁹, —NR⁵⁹SO₂R⁵⁹, OH, CN,

—CO₂H,

—R⁵⁹—OH, —O—R⁵⁹, —COOR¹⁹,

—CON(R⁵⁹)₂, —CONROR⁵⁹,

—SO₃H, —R⁵⁹—S, —SO₂N(R⁵⁹)₂,

—S(O)R⁵⁹, —S(O)₂R⁵⁹

wherein each R⁵⁹ is independently an aryl or aliphatic, optionally withsubstitution. Among hetero substituents containing R⁵⁹ groups,preference is given to those materials having aryl and alkyl R⁵⁹ groupsas defined herein. Preferred hetero substituents are those listed above.

“Hydrogen bond donor” group refers to a group containg O—H, or N—Hfunctionality. Examples of “hydrogen bond donor” groups include —OH,—NH₂, and —NH—R^(59a) and wherein R^(59a) is alkyl, acyl, cycloalkyl,aryl, or heteroaryl.

“Dihydroxyphosphoryl” refers to the radical —PO(OH)₂.

“Substituted dihydroxyphosphoryl” includes those groups recited in thedefinition of “substituted” herein, and particularly refers to adihydroxyphosphoryl radical wherein one or both of the hydroxyl groupsare substituted. Suitable substituents are described in detail below.

“Aminohydroxyphosphoryl” refers to the radical —PO(OH)NH₂.

“Substituted aminohydroxyphosphoryl” includes those groups recited inthe definition of “substituted” herein, and particularly refers to anaminohydroxyphosphoryl wherein the amino group is substituted with oneor two substituents. Suitable substituents are described in detailbelow. In certain embodiments, the hydroxyl group can also besubstituted.

“Thioalkoxy” refers to the group —SR⁶⁰ where R⁶⁰ is alkyl.

“Substituted thioalkoxy” includes those groups recited in the definitionof “substituted” herein, and particularly refers to a thioalkoxy grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, selected from the groupconsisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino,aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy,azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen,hydroxyl, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy,thioketo, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— and aryl-S(O)₂—.

“Sulfanyl” refers to the radical HS—. “Substituted sulfanyl” refers to aradical such as RS— wherein R is any substituent described herein.

“Sulfonyl” refers to the divalent radical —S(O₂)—. “Substitutedsulfonyl” refers to a radical such as R⁶¹—(O₂)S— wherein R⁶¹ is anysubstituent described herein. “Aminosulfonyl” or “Sulfonamide” refers tothe radical H₂N(O₂)S—, and “substituted aminosulfonyl” or “substitutedsulfonamide” refers to a radical such as R⁶² ₂N(O₂)S— wherein each R⁶²is independently any substituent described herein.

“Sulfone” refers to the group —SO₂R⁶³. In particular embodiments, R⁶³ isselected from H, lower alkyl, alkyl, aryl and heteroaryl.

“Thioaryloxy” refers to the group —SR⁶⁴ where R⁶⁴ is aryl.

“Thioketo” refers to the group ═S.

“Thiol” refers to the group —SH.

One having ordinary skill in the art of organic synthesis will recognizethat the maximum number of heteroatoms in a stable, chemically feasibleheterocyclic ring, whether it is aromatic or non aromatic, is determinedby the size of the ring, the degree of unsaturation and the valence ofthe heteroatoms. In general, a heterocyclic ring may have one to fourheteroatoms so long as the heteroaromatic ring is chemically feasibleand stable.

“Pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopoeia orother generally recognized pharmacopoeia for use in animals, and moreparticularly in humans.

“Pharmaceutically acceptable salt” refers to a salt of a compoundprovided herein that is pharmaceutically acceptable and that possessesthe desired pharmacological activity of the parent compound. Such saltsinclude: (1) acid addition salts, formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or formed with organic acids such asacetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid,glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid,malic acid, maleic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelicacid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, N-methylglucamine and thelike. Salts further include, by way of example only, sodium, potassium,calcium, magnesium, ammonium, tetraalkylammonium, and the like; and whenthe compound contains a basic functionality, salts of non toxic organicor inorganic acids, such as hydrochloride, hydrobromide, tartrate,mesylate, acetate, maleate, oxalate and the like. The term“pharmaceutically acceptable cation” refers to a non toxic, acceptablecationic counter-ion of an acidic functional group. Such cations areexemplified by sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium cations, and the like.

“Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant,excipient or carrier with which a compound provided herein isadministered.

‘Preventing’ or ‘prevention’ refers to a reduction in risk of acquiringor developing a disease or disorder (i.e., causing at least one of theclinical symptoms of the disease not to develop in a subject that may beexposed to a disease-causing agent, or predisposed to the disease inadvance of disease onset.

“Prodrugs” refers to compounds, including derivatives of the compoundsprovided herein, which have cleavable groups and become by solvolysis orunder physiological conditions the compounds provided herein which arepharmaceutically active in vivo. Such examples include, but are notlimited to, choline ester derivatives and the like, N-alkylmorpholineesters and the like.

The term ‘prophylaxis’ is related to ‘prevention’, and refers to ameasure or procedure the purpose of which is to prevent, rather than totreat or cure a disease. Non-limiting examples of prophylactic measuresmay include the administration of vaccines; the administration of lowmolecular weight heparin to hospital patients at risk for thrombosisdue, for example, to immobilization; and the administration of ananti-malarial agent such as chloroquine, in advance of a visit to ageographical region where malaria is endemic or the risk of contractingmalaria is high.

“Solvate” refers to forms of the compound that are associated with asolvent, usually by a solvolysis reaction. Conventional solvents includewater, ethanol, acetic acid and the like. The compounds provided hereinmay be prepared e.g. in crystalline form and may be solvated orhydrated. Suitable solvates include pharmaceutically acceptablesolvates, such as hydrates, and further include both stoichiometricsolvates and non-stoichiometric solvates.

“Subject” includes humans. The terms “human,” “patient” and “subject”are used interchangeably herein.

“Tautomers” refer to compounds that are interchangeable forms of aparticular compound structure, and that vary in the displacement ofhydrogen atoms and electrons. Thus, two structures may be in equilibriumthrough the movement of π electrons and an atom (usually H). Forexample, enols and ketones are tautomers because they are rapidlyinterconverted by treatment with either acid or base. Another example oftautomerism is the aci- and nitro-forms of phenylnitromethane, that arelikewise formed by treatment with acid or base.

Tautomeric forms may be relevant to the attainment of the optimalchemical reactivity and biological activity of a compound of interest.

“Therapeutically effective amount” means the amount of a compound that,when administered to a subject for treating a disease, is sufficient toeffect such treatment for the disease. The “therapeutically effectiveamount” can vary depending on the compound, the disease and itsseverity, and the age, weight, etc., of the subject to be treated.

“Treating” or “treatment” of any disease or disorder refers, in oneembodiment, to ameliorating the disease or disorder (i.e., arresting orreducing the development of the disease or at least one of the clinicalsymptoms thereof). In another embodiment “treating” or “treatment”refers to ameliorating at least one physical parameter, which may not bediscernible by the subject. In yet another embodiment, “treating” or“treatment” refers to modulating the disease or disorder, eitherphysically, (e.g., stabilization of a discernible symptom),physiologically, (e.g., stabilization of a physical parameter), or both.In yet another embodiment, “treating” or “treatment” refers to delayingthe onset of the disease or disorder.

Other derivatives of the compounds provided herein have activity in boththeir acid and acid derivative forms, but in the acid sensitive formoften offers advantages of solubility, tissue compatibility, or delayedrelease in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well know to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides and anhydridesderived from acidic groups pendant on the compounds provided herein arepreferred prodrugs. In some cases it is desirable to prepare doubleester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. Preferred are the C₁ to C₈ alkyl,C₂-C₈ alkenyl, aryl, C₇-C₁₂ substituted aryl, and C₇-C₁₂ arylalkylesters of the compounds provided herein.

As used herein, the term “isotopic variant” refers to a compound thatcomprises an unnatural proportion of an isotope of one or more of theatoms that constitute such compound. For example, an “isotopic variant”of a compound can comprise an unnatural proportion of one or morenon-radioactive isotopes, such as for example, deuterium (²H or D),carbon-13 (¹³C), nitrogen-15 (¹⁵N), or the like. It will be understoodthat, in a compound comprising an unnatural proportion of an isotope,any example of an atom where present, may vary in isotope composition.For example, any hydrogen may be ²H/D, or any carbon may be ¹³C, or anynitrogen may be ¹⁵N, and that the presence and placement of such atomsmay be determined within the skill of the art. Likewise, provided hereinare methods for preparation of isotopic variants with radioisotopes, inthe instance for example, where the resulting compounds may be used fordrug and/or substrate tissue distribution studies. The radioactiveisotopes tritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularlyuseful for this purpose in view of their ease of incorporation and readymeans of detection. Further, compounds may be prepared that aresubstituted with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, and would be useful in Positron Emission Topography (PET) studiesfor examining substrate receptor occupancy. All isotopic variants of thecompounds provided herein, radioactive or not, are intended to beencompassed within the scope provided herein.

It is also to be understood that compounds that have the same molecularformula but differ in the nature or sequence of bonding of their atomsor the arrangement of their atoms in space are termed “isomers”. Isomersthat differ in the arrangement of their atoms in space are termed“stereoisomers”.

Stereoisomers that are not mirror images of one another are termed“diastereomers” and those that are non-superimposable mirror images ofeach other are termed “enantiomers”. When a compound has an asymmetriccenter, for example, it is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center and is described by theR- and S-sequencing rules of Cahn and Prelog, or by the manner in whichthe molecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a “racemic mixture”.

As used herein a pure enantiomeric compound is substantially free fromother enantiomers or stereoisomers of the compound (i.e., inenantiomeric excess). In other words, an “S” form of the compound issubstantially free from the “R” form of the compound and is, thus, inenantiomeric excess of the “R” form. The term “enantiomerically pure” or“pure enantiomer” denotes that the compound comprises more than 75% byweight, more than 80% by weight, more than 85% by weight, more than 90%by weight, more than 91% by weight, more than 92% by weight, more than93% by weight, more than 94% by weight, more than 95% by weight, morethan 96% by weight, more than 97% by weight, more than 98% by weight,more than 98.5% by weight, more than 99% by weight, more than 99.2% byweight, more than 99.5% by weight, more than 99.6% by weight, more than99.7% by weight, more than 99.8% by weight or more than 99.9% by weight,of the enantiomer. In certain embodiments, the weights are based upontotal weight of all enantiomers or stereoisomers of the compound.

As used herein and unless otherwise indicated, the term“enantiomerically pure R-compound” refers to at least about 80% byweight R-compound and at most about 20% by weight S-compound, at leastabout 90% by weight R-compound and at most about 10% by weightS-compound, at least about 95% by weight R-compound and at most about 5%by weight S-compound, at least about 99% by weight R-compound and atmost about 1% by weight S-compound, at least about 99.9% by weightR-compound or at most about 0.1% by weight S-compound. In certainembodiments, the weights are based upon total weight of compound.

As used herein and unless otherwise indicated, the term“enantiomerically pure S-compound” or “S-compound” refers to at leastabout 80% by weight S-compound and at most about 20% by weightR-compound, at least about 90% by weight S-compound and at most about10% by weight R-compound, at least about 95% by weight S-compound and atmost about 5% by weight R-compound, at least about 99% by weightS-compound and at most about 1% by weight R-compound or at least about99.9% by weight S-compound and at most about 0.1% by weight R-compound.In certain embodiments, the weights are based upon total weight ofcompound.

In the compositions provided herein, an enantiomerically pure compoundor a pharmaceutically acceptable salt, solvate, hydrate or prodrugthereof can be present with other active or inactive ingredients. Forexample, a pharmaceutical composition comprising enantiomerically pureR-compound can comprise, for example, about 90% excipient and about 10%enantiomerically pure R-compound. In certain embodiments, theenantiomerically pure R-compound in such compositions can, for example,comprise, at least about 95% by weight R-compound and at most about 5%by weight S-compound, by total weight of the compound. For example, apharmaceutical composition comprising enantiomerically pure S-compoundcan comprise, for example, about 90% excipient and about 10%enantiomerically pure S-compound. In certain embodiments, theenantiomerically pure S-compound in such compositions can, for example,comprise, at least about 95% by weight S-compound and at most about 5%by weight R-compound, by total weight of the compound. In certainembodiments, the active ingredient can be formulated with little or noexcipient or carrier.

The compounds provided herein may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers andmixtures, racemic or otherwise, thereof. The methods for thedetermination of stereochemistry and the separation of stereoisomers arewell-known in the art.

The S-Triazine Compounds

As set forth earlier herein, the s-triazine compounds compriseantimicrobial/antiviral/antifungal s-triazine compounds. The s-triazinecompounds may have a lethal effect on bacteria, viruses or fungi in itsmonomeric form. More particularly, the s-triazine compounds may be anyantimicrobial s-triazine compounds, including natural products found inorganisms, fragments of natural s-triazine compounds, and any syntheticanalogs or de novo designs. These s-triazine compounds can accordinglyinclude normatural amino acids: beta-amino acids, d-amino acids and/ornon-indigenous amino acids.

The present invention relates to s-triazine compounds havingantimicrobial properties, according to formula I:

wherein

each R¹, R^(1a) and R² is independently substituted amino; or apharmaceutically acceptable salt, solvate or prodrug thereof; andstereoisomers, isotopic variants and tautomers thereof.

More particularly, the present invention relates to s-triazine compoundshaving antimicrobial properties, according to formula Ia:

wherein

-   -   each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f), R^(2g),        R^(2h) and R^(2j) is independently selected from hydrogen, and        substituted or unsubstituted C₁-C₆ alkyl; or any two of R^(2c),        R^(2d), R^(2e), R^(2f) and R^(2g) may join together to form a        cycloalkyl or cycloheteroalkyl ring or 3-7 atoms;    -   R³ is R⁴ or substituted or unsubstituted C₁-C₆ alkyl; or is        selected from a 3-13 membered cycloalkyl, heterocycloalkyl, aryl        and heteroaryl ring system, which can be optionally substituted        with one or more substituents independently selected from halo,        hydroxyl, amino, cyano, sulfo, sulfanyl, sulfinyl, amido,        carboxy, ester, alkyl, substituted alkyl, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, and sulfonamido;    -   R⁴ is

-   -   wherein C¹, C² and C³ taken together with the C atom to which        they are attached to form a bi- or tri-cyloalkyl or        cycloheteroalkyl ring system of 7-13 atoms; and wherein the ring        system is substituted or unsubstituted;    -   m is 0, 1, 2 or 3; and n is 2, 3 or 4;    -   or a pharmaceutically acceptable salt, solvate or prodrug        thereof;    -   and stereoisomers, isotopic variants and tautomers thereof.

In one embodiment, with respect to compounds of formula Ia, R⁴ issubstituted or unsubstituted

In one embodiment, with respect to compounds of formula I, R⁴ is

-   -   and wherein R^(a), R^(b) and R^(c) are independently selected        from H, halo, hydroxyl, alkyl, amino, and aryl.

In one embodiment, with respect to compounds of formula Ia, R⁴ is asdescribed above and each R^(a), R^(b) and R^(c) is independentlyselected from H, Br, Cl, OH, Me, NHAc, Ph and F. In a preferredembodiment, each R^(a), R^(b) and R^(c) is H.

In one embodiment, with respect to compounds of formula Ia, each R^(2a),R^(2b), R^(2c), R^(2d), R^(2e), R^(2f), R^(2g), R^(2h) and R^(2j) isindependently hydrogen.

In one embodiment, with respect to compounds of formula Ia, one ofR^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f), R^(2g), R^(2h) andR^(2j) is substituted or unsubstituted C₁-C₆ alkyl and the rest arehydrogen.

In one embodiment, with respect to compounds of formula Ia, one ofR^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f), R^(2g), R^(2h) andR^(2j) is methyl and the rest are hydrogen.

In one embodiment, with respect to compounds of formula Ia, each ofR^(2c), R^(2d), and R^(2f) is hydrogen and R^(2e), and R^(2g) are joinedtogether to form a 4-7 membered heterocycloalkyl ring.

In one embodiment, with respect to compounds of formula Ia, R^(2b) issubstituted or unsubstituted C₁-C₆ alkyl.

In one embodiment, with respect to compounds of formula Ia, R^(2b) ismethyl, ethyl, or propyl.

Another aspect of invention provides a s-triazine compound having aformula:

-   -   wherein    -   each of R^(2f), and R^(2g) is independently selected from        hydrogen, and substituted or unsubstituted C₁-C₆ alkyl;    -   R³ is R⁴ or substituted or unsubstituted C₁-C₆ alkyl; or is        selected from a 3-13 membered cycloalkyl, heterocycloalkyl, aryl        and heteroaryl ring system, which can be optionally substituted        with one or more substituents independently selected from halo,        hydroxyl, amino, cyano, sulfo, sulfanyl, sulfinyl, amido,        carboxy, ester, alkyl, substituted alkyl, alkenyl, substituted        alkenyl, alkynyl, substituted alkynyl, and sulfonamido;    -   m is 0, 1, 2 or 3; and n is 2, 3 or 4;    -   or a pharmaceutically acceptable salt, solvate or prodrug        thereof;    -   and stereoisomers, isotopic variants and tautomers thereof.

In one embodiment, with respect to compounds of formula II, n is 2, 3 or4.

In one embodiment, with respect to compounds of formula II, n is 2 or 3.

In one embodiment, with respect to compounds of formula II, n is 3.

In one embodiment, with respect to compounds of formula II, each ofR^(2f) and R^(2g) is independently selected from hydrogen, andsubstituted or unsubstituted C₁-C₆ alkyl.

In one embodiment, with respect to compounds of formula II, each ofR^(2f) and R^(2g) is hydrogen.

In one embodiment, with respect to compounds of formula II, one ofR^(2f) and R^(2g) is hydrogen, and the other is substituted orunsubstituted C₁-C₆ alkyl.

In one embodiment, with respect to compounds of formula II, one ofR^(2f) and R^(2g) is hydrogen, and the other is methyl, ethyl, n-propylor n-butyl.

In one embodiment, with respect to compounds of formula II, each ofR^(2f) and R^(2g) is independently substituted or unsubstituted C₁-C₆alkyl.

In one embodiment, with respect to compounds of formula II, each ofR^(2f), and R^(2g) is independently methyl, ethyl, n-propyl or n-butyl.

In one embodiment, with respect to compounds of formula II, each ofR^(2f) and R^(2g) is methyl, ethyl, n-propyl or n-butyl.

In one embodiment, with respect to compounds of formula II, each ofR^(2f) and R^(2g) is n-butyl.

In one embodiment, with respect to compounds of formula II, wherein m is0.

In one embodiment, with respect to compounds of formula II, wherein m is1.

In one embodiment, with respect to compounds of formula II, wherein m is2.

In one embodiment, with respect to compounds of formula II, wherein m is3.

In one embodiment, with respect to compounds of formula II, wherein R³is selected from a 3-13 membered cycloalkyl, heterocycloalkyl, aryl andheteroaryl ring system, which can be optionally substituted with one ormore substituents independently selected from halo, hydroxyl, amino,cyano, sulfo, sulfanyl, sulfinyl, amido, carboxy, ester, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, and sulfonamide.

In one embodiment, with respect to compounds of formula II, R³ isselected from 3-13 membered cycloalkyl ring.

In one embodiment, with respect to compounds of formula II, R³ iscyclopropyl or cyclobutyl.

In one embodiment, with respect to compounds of formula II, R³ iscyclopentyl or cyclohexyl.

In one embodiment, with respect to compounds of formula II, R³ iscycloheptyl.

In one embodiment, with respect to compounds of formula II, R³ is R⁴ andR⁴ is as described for formula Ia.

In one embodiment, with respect to compounds of formula II, R³ is R⁴ andR⁴ is selected from substituted or unsubstituted

In one embodiment, with respect to compounds of formula II, R³ isadamantane ring.

In one embodiment, with respect to compounds of formula II, R³ isnorbornane ring.

In one embodiment, with respect to compounds of formula II, R³ issubstituted or unsubstituted C₁-C₆ alkyl.

In one embodiment, with respect to compounds of formula II, R³ ismethyl, ethyl, n-propyl or n-butyl.

In one embodiment, with respect to compounds of formula II, R³ isselected from a 6-13 membered aryl and heteroaryl ring system, which canbe optionally substituted with one or more substituents independentlyselected from halo, hydroxyl, amino, cyano, sulfo, sulfanyl, sulfinyl,amido, carboxy, ester, alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, and sulfonamide.

In one embodiment, with respect to compounds of formula II, R³ isphenyl, which can be optionally substituted with one or moresubstituents independently selected from halo, hydroxyl, amino, cyano,sulfo, sulfanyl, sulfinyl, amido, carboxy, ester, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, andsulfonamide.

In one embodiment, with respect to compounds of formula II, R³ isunsubstituted phenyl.

In one embodiment, with respect to compounds of formula II, the compoundis a quaternary salt.

In one embodiment, with respect to compounds of formula II, the compoundis selected from:

In one embodiment, with respect to compounds of formula II, the compoundis:

In certain aspects and where appropriate, the present invention extendsto the preparation of prodrugs and derivatives of the compounds of theinvention. Prodrugs are derivatives which have cleavable groups andbecome by solvolysis or under physiological conditions the compounds ofthe invention, which are pharmaceutically active, in vivo.

Pharmaceutical Compositions

When employed as pharmaceuticals, the triazine compounds of thisinvention are typically administered in the form of a pharmaceuticalcomposition. Such compositions can be prepared in a manner well known inthe pharmaceutical art and comprise at least one active compound.

Generally, the triazine compounds of this invention is administered in apharmaceutically effective amount. The amount of the compound actuallyadministered will typically be determined by a physician, in the lightof the relevant circumstances, including the condition to be treated,the chosen route of administration, the actual compound administered,the age, weight, and response of the individual patient, the severity ofthe patient's symptoms, and the like.

The pharmaceutical compositions of this invention can be administered bya variety of routes including by way of non limiting example, oral,rectal, transdermal, subcutaneous, intravenous, intramuscular andintranasal. Depending upon the intended route of delivery, the compoundsof this invention are preferably formulated as either injectable or oralcompositions or as salves, as lotions or as patches all for transdermaladministration.

The compositions for oral administration can take the form of bulkliquid solutions or suspensions, or bulk powders. More commonly,however, the compositions are presented in unit dosage forms tofacilitate accurate dosing. The term “unit dosage forms” refers tophysically discrete units suitable as unitary dosages for human subjectsand other mammals, each unit containing a predetermined quantity ofactive material calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient. Typical unitdosage forms include prefilled, premeasured ampoules or syringes of theliquid compositions or pills, tablets, capsules or the like in the caseof solid compositions. In such compositions, the furansulfonic acidcompound is usually a minor component (from about 0.1 to about 50% byweight or preferably from about 1 to about 40% by weight) with theremainder being various vehicles or carriers and processing aids helpfulfor forming the desired dosing form.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like. Solid forms may include, forexample, any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art. As before, the active compound in such compositions istypically a minor component, often being from about 0.05 to 10% byweight with the remainder being the injectable carrier and the like.

Transdermal compositions are typically formulated as a topical ointmentor cream containing the active ingredient(s), generally in an amountranging from about 0.01 to about 20% by weight, preferably from about0.1 to about 20% by weight, preferably from about 0.1 to about 10% byweight, and more preferably from about 0.5 to about 15% by weight. Whenformulated as an ointment, the active ingredients will typically becombined with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream with,for example an oil-in-water cream base. Such transdermal formulationsare well-known in the art and generally include additional ingredientsto enhance the dermal penetration of stability of the active ingredientsor the formulation. All such known transdermal formulations andingredients are included within the scope of this invention.

The compounds and compositions of the invention can also be administeredby a transdermal device. Accordingly, transdermal administration can beaccomplished using a patch either of the reservoir or porous membranetype, or of a solid matrix variety.

The above-described components for orally administrable, injectable ortopically administrable compositions are merely representative. Othermaterials as well as processing techniques and the like are set forth inPart 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, MackPublishing Company, Easton, Pa., which is incorporated herein byreference.

The compounds and compositions of the invention can also be administeredin sustained release forms or from sustained release drug deliverysystems. A description of representative sustained release materials canbe found in Remington's Pharmaceutical Sciences.

The following formulation examples illustrate representativepharmaceutical compositions that may be prepared in accordance with thisinvention. The present invention, however, is not limited to thefollowing pharmaceutical compositions.

Formulation 1—Tablets

A compound of the invention is admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into240-270 mg tablets (80-90 mg of active compound per tablet) in a tabletpress.

Formulation 2—Capsules

A compound of the invention is admixed as a dry powder with a starchdiluent in an approximate 1:1 weight ratio. The mixture is filled into250 mg capsules (125 mg of active compound per capsule).

Formulation 3—Liquid

A compound of the invention (125 mg) may be admixed with sucrose (1.75g) and xanthan gum (4 mg) and the resultant mixture may be blended,passed through a No. 10 mesh U.S. sieve, and then mixed with apreviously made solution of microcrystalline cellulose and sodiumcarboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10mg), flavor, and color are diluted with water and added with stirring.Sufficient water may then added to produce a total volume of 5 mL.

Formulation 4—Tablets

A compound of the invention may be admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into450-900 mg tablets (150-300 mg of active compound) in a tablet press.

Formulation 5—Injection

A compound of the invention is dissolved or suspended in a bufferedsterile saline injectable aqueous medium to a concentration ofapproximately 5 mg/mL.

Formulation 6—Topical

Stearyl alcohol (250 g) and a white petrolatum (250 g) are melted atabout 75° C. and then a mixture of a compound of the invention (50 g)methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate(10 g), and propylene glycol (120 g) dissolved in water (about 370 g) isadded and the resulting mixture is stirred until it congeals.

Methods of Treatment

The present compounds may be used as therapeutic agents for thetreatment of conditions in mammals. Accordingly, the compounds andpharmaceutical compositions of this invention find use as therapeuticsfor preventing and/or treating infections and like maladies resultingfrom bacterial, viral or fungal attack, and related conditions inmammals including humans.

In a method of treatment aspect, this invention provides a method oftreating a mammal susceptible to or afflicted with a conditionassociated with or resulting from bacterial, viral or fungal attack orinfection, which method comprises administering an effective amount ofone or more of the pharmaceutical compositions just described.

In additional method of treatment aspects, this invention providesmethods of treating a mammal susceptible to or afflicted with a varietyof bacteria or other infections, including strains which have developedresistance to traditional antibiotics, such as, for example,Staphylococcus aureus. The method comprises administering an effectivecondition-treating or condition-preventing amount of one or more of thepharmaceutical compositions just described.

Injection dose levels range from about 0.1 mg/kg/hour to at least 10mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kgor more may also be administered to achieve adequate steady statelevels. The maximum total dose is not expected to exceed about 2 g/dayfor a 40 to 80 kg human patient.

For the prevention and/or treatment of long-term conditions, such asviral or microbial conditions, the regimen for treatment usuallystretches over many months or years so oral dosing is preferred forpatient convenience and tolerance. With oral dosing, one to five andespecially two to four and typically three oral doses per day arerepresentative regimens. Using these dosing patterns, each dose providesfrom about 0.01 to about 20 mg/kg of the compound or its derivative,with preferred doses each providing from about 0.1 to about 10 mg/kg andespecially about 1 to about 5 mg/kg.

Transdermal doses are generally selected to provide similar or lowerblood levels than are achieved using injection doses.

The compounds of this invention can be administered as the sole activeagent or they can be administered in combination with other agents,including other active derivatives.

General Synthetic Procedures

The compounds of this invention can be prepared from readily availablestarting materials using the general methods and procedures describedearlier and illustrated schematically in the examples that follow. Itwill be appreciated that where typical or preferred process conditions(i.e., reaction temperatures, times, mole ratios of reactants, solvents,pressures, etc.) are given, other process conditions can also be usedunless otherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvent used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and P. G. M. Wuts, ProtectingGroups in Organic Synthesis, Second Edition, Wiley, New York, 1991, andreferences cited therein.

The following methods are presented with details as to the preparationof representative s-triazine compounds that have been listedhereinabove. The s-triazine compounds of the invention may be preparedfrom known or commercially available starting materials and reagents byone skilled in the art of organic synthesis.

With respect to the Figures and Schemes appearing below, it should beunderstood that reference to R² pertains to the below Figures andSchemes, and that reference to R^(2a)-R^(2j) appearing elsewhere herein,pertains to individual substituents that may be attached to N andpositioned, among other places, at the location of R2. Naturally, theplacement of R^(2a)-R^(2j) may vary within the scope and contemplationof the invention.

General

Unless otherwise noted, materials and solvents were obtained fromcommercial suppliers and were used without further purification.Anhydrous tetrahydrofuran (THF) and 1-methyl-2-pyrrolidinone (NMP) fromAcros were used as reaction solvents without any prior purification.PAL-aldehyde resin from Midwest Bio-Tech was used as the solid support.For the synthesis of building block I, general coupling reactions wereperformed through solution phase chemistry and were purified by flashcolumn chromatography on Merck silica gel 60-PF₂₄₅. All products wereidentified by LC/MS from Agilent Technology using a C18 column (20×4.0mm), with a gradient of 5-95% CH₃CN (containing 1% acetic acid)-H₂O(containing 1% acetic acid) as eluant.

Equipment

Thermal reactions were performed using a standard heat block from VWRscientific Products using 4 mL vials. Resin filtration procedures werecarried out using 70μ PE flit cartridge from Applied Separations.

General schemes for the preparation of Libriaries 1 to 5 are set forthdiagrammatically below.

While the above schemes provide an overview of the synthetic pathways,the following provides the details of these procedures.

Synthesis of Triazine Libraries

Synthesis of the TG-Boc Linker (1) for Library 2

2,2′-(Ethylenedioxy)bis(ethylamine) (10 equiv.) was dissolved indichloromethane and the solution was cooled down to −78° C. in a dryice/acetone bath. Di-tert-butyl dicarbonate (1 equiv.) was dissolved indichloromethane and added to the solution of2,2′-(Ethylenedioxy)bis(ethylamine) dropwise over a period of 3 h in anitrogen gas atmosphere. The reaction mixture was allowed to stir for 10h followed by extraction with saturated NaCl solution. The organiclayers were combined and dried over MgSO₄. The solvent was removed invacuo (70% yield).

General Procedure for Preparing Building Block II via GrignardAlkylation for Library 2

A solution with anhydrous THF (125 mL) of alkylmagnesiumhalide(Br/Cl)(13.0 mmol) was slowly added to a cooled (0° C.-5° C.), mechanicallystirred THF solution (125 mL) of cyanuric chloride (10.93 mmol, 2 g);the mixture was stirred at 0° C. for 2 h. The reaction mixture wasquenched with 1 N HCl (40 mL). Then the reaction mixture was extractedwith ethyl acetate and washed with water. The organic layers werecombined and dried over MgSO₄. The solvent was removed in vacuo.

General Procedure for Building Block I Synthesis for Library 1 to 5

2.8 mmole (5 equiv) of R₁ amines (in library 1, 3, 4 and 5) or TG-Boclinker (for library 2).) was added to a suspension of the PAL aldehyderesin (1.3 g, 1.43 mmole) in anhydrous tetrahydrofuran (THF) (50 mLcontaining 2% of acetic acid) at room temperature. The reaction mixturewas shaken for 1 h at room temperature followed by addition of sodiumtriacetoxyborohydride (2.1 g, 9.9 mmole, 7 equiv.). The reaction mixturewas stirred for 12 h and filtered. The resin was washed withN,N-dimethylformamide (DMF) (5 times), alternatively withdichloromethane and methanol (MeOH) (5 times), and finally withdichloromethane (5 times). The resin was dried in vacuo.

Synthesis of Building Blocks II by Solution Phase Synthesis for Library1, 3, 4 and 5

Cyanuric trichloride (1 equiv.) was dissolved in THF with DIEA (10equiv.) at 0° C. 1-Adamantylamine (1.2 equiv.) in THF was addeddropwise. The reaction mixture was stirred at 0° C. for 1 h. A solidprecipitate slowly formed. Upon completion of the reaction, the reactionmixture was quickly filtered through a plug of flash silica and washedwith EA. The filtrate was evaporated in vacuo. The resulting productswere purified using flash column chromatography (particle size 32-63μm).

General Procedure for Coupling Building Block I and Building Block II

Building Block II (0.44 mmole) was added to Building Block I (0.11mmole) in DIEA (1 mL) and anhydrous THF (10 mL) at room temperature. Thereaction mixture was heated to 60° C. for 3 h and filtered. The resinwas washed with DMF (5 times), alternatively with dichloromethane andmethanol (5 times), and finally dichloromethane (5 times). The resin wasdried in vacuo.

General Procedure for the Final Amination on the Resin and ProductCleavage Reaction.

The desired amines (4 equiv.) were added to the resin (10 mg), coupledwith Building Block I and Building Block II, in DIEA (8 μL) and 1 mL ofNMP: n-BuOH (1:1). The reaction mixture was heated to 120° C. for 3 h.The resin was washed with DMF (5 times), alternatively withdichloromethane and methanol (5 times), and finally dichloromethane (5times). The resin was dried in vacuo. The product cleavage reaction wasperformed using 10% trifluoroacetic acid (TFA) in dichloromethane (1 mL)for 30 min at room temperature and washed with dichloromethane (0.5 mL).Free hydroxyl containing compounds were further treated with apiperazine resin in 0.5 mL THF at room temperature for 5 h to cleave thetrifluoroacetic ester that was formed upon treatment with TFA. The resinwas filtered out and washed with 0.1 mL THF. The purity and identity ofall the products were monitored by LC-MS at 250 nm (Agilent Model 1100)and more than 90% of the compounds demonstrated >90% purity.

The first two libraries were created in order to screen variousfunctional groups for their antimicrobial activity. Two mono-substitutedtriazine templates were used for these libraries, where amantadine and2,2′-(Ethylenedioxy)bis(ethylamine) were prefixed onto the triazinescaffold (FIG. 1). Amantadine was used to represent one hydrophobicgroup, and 2,2′-(Ethylenedioxy)bis(ethylamine) was used to represent onecharged group.

In addition of the prefixed group, the series of compounds were made tobring in two more groups (hydrophobic, bulky, positively charged, etc)to positions R¹ and R², respectively. A total of 80 compounds in Library1 and 72 compounds in Library 2 were synthesized utilizing an orthogonalsynthetic approach based on a triazine scaffold (Scheme 1) and theirantimicrobial activities were tested against gram-positive bacteria B.Subtilis. The subsequent Libraries 3 and 4 were designed based on theresults from libraries 1 and 2 (FIG. 2 and Scheme 2), and Library 5 wasmade to further optimize the R¹ and R² groups of the most promisingcompounds from previous design (FIG. 3 and Scheme 2).

I

For Library 1, R^(1a) is —NH-adamantane; and For Library 2, R^(1a) is—NH(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂NH₂ R1 Structure R2 Structure A

1

B

2

C

3

D

4

E

5

F

6

G

7

H

8

I             J

A

1

B

2

C

3

D

4

E

5

F

6

G

7

H

8

9

I

For Library 3, R^(1a) is —NH-adamantane; and For Library 4, R^(1a) is—NH-adamantane; and R² is —NH(CH₂)₂—O—(CH₂)₂—O—(CH₂)₂NH₂ R1 Structure R2Structure A

A

B

B

1         2

3

4

5

6

7

R1 Structure

 1        2

 3

 4

 5

 6

 7

 8

 9

10

11

EXAMPLES

The following examples are presented in order to more fully illustratethe preferred embodiments of the invention. They should in no way beconstrued, however, as limiting the broad scope of the invention.

Example 1

Antibacterial Testing

The antimicrobial activity of each triazine compound was tested byfollowing standard broth microdilution protocols recommended by theNational Committee for Clinical Laboratory Standard. All compounds wereinitially tested against Bacillus subtilis (ATCC 6633; Rockville, Md.).The three compounds showing the most antimicrobial activity againstBacillus subtilis, A7 (Library 3), A4 (Library 5) and C2 (Library 5),were further tested against ampicillin- and streptomycin-resistantstrain E. coli (D31; E. coli Genetic Resource Center, Yale University,New Haven, Conn.), multi-drug resistant strain S. aureus (ATCC BAA-44),Acinetobacter baumannii (ATCC BAA-747), and Enterococcus faecalis (ATCC29212). Bacteria were grown in Mueller Hinton Broth (MHB) at 37° C. forovernight. Then, cultures were diluted in MHB to a final concentrationof 2×10⁴ to 2×10⁵ CFU/mL. Bacterial inocula were incubated at 37° C. inPBS buffer (pH 7.2) with varying concentrations of 2-fold dilution oftriazine stocks. The 18-hour absorbance data were used to calculate thepercent inhibition for each sample by comparing with the absorbance ofcultures without triazine compounds. Bacterial growth was measured byturbidity as optical density at 600 nm using a Genesys 5Spectrophotometer (Rochester, N.Y.). The concentration of peptide thatresulted in 50% was recorded as MIC₅₀.

Example 2

Hemolytic Activity

Hemolytic activity of three triazine compounds (A7 from library 3, A4and C2 from library 5) was assessed on fresh sheep erythrocytes(Fitzgerald Inc., Concord, Mass.). The red blood cell suspension wasincubated in PBS buffer (pH 7.2) with varying concentrations of samplestocks at 37° C. for 30 minutes, and then spun down at 3,000 rpm for 10mins. The resulting supernatant was diluted by a factor of 40 indistilled water. The absorbance of the supernatant at 540 nm wasmeasured using a Genesys 5 Spectrophotometer. Zero hemolysis and 100%hemolysis controls were obtained by incubating the cells with buffer and1% Triton-X, respectively. Sample concentration yielding 50% hemolysiswas used as hemolytic dose (HD₅₀) determined from dose-response curves.

Results

The initial screening of two groups of mono-substituted 1,3,5-triazinecompounds (Libraries 1 & 2) against gram-positive bacteria B. subtilisat 100 uM concentration revealed 30 antimicrobial active compounds.However, none of these compounds showed antimicrobial activity at lowerconcentration (50 uM). In order to improve the activity, the R¹ and R²side chains in each of the 30 compounds were organized and tables offrequency of effectiveness from Libraries 1 and 2 were made (Tables 1and 2). From these tables, most effective side chains from the twolibraries were identified, and were then used to design Libraries 3 and4. Due to synthesis difficulties with some of the compounds, onlylimited combinations of these groups were synthesized.

TABLE 1 Frequency of effectiveness from library 1 (active against B.sub. at 100 uM;: B2, C2, F2, H2, F3, G3, H3, E6, C7, H7, C8, F8 G8). I

R1 Structure Freq. A

0 B

1 C

3 D

0 E

1 F

3 G

2 H

3 I

0 J

0 R2 Structure Freq. 1

0 2

4 3

3 4

0 5

0 6

1 7

2 8

3

TABLE 2 Frequency of effectiveness from library 2 (active against B.sub. at 100 uM: B1, C1, E1, F1, C3, E3, F3, C5, E5, F5, E6,; A7, B7, C7,E7, F7, H7). I

R1 Structure Freq. A

1 B

2 C

4 D

0 E

5 F

4 G

0 H

1 R2 Structure Freq. 1

4 2

0 3

3 4

0 5

3 6

1 7

6 8

0 9

0

The screening of compounds in Libraries 3 & 4 against Gram-positivebacteria B. subtilis further revealed 11 compounds that had improvedantimicrobial activities, and their hemolytic activities (Table 3).Among these compounds, A7 and B4 from Library 3 were selected as thetemplates for designing Library 5. Various length, charge, or bulkinesswere tested in Library 5 in order to optimize the compounds, as well asto gain further insight as to the structure-function relationship ofthese peptide mimics.

TABLE 3 Successful screening results from library 3 & 4 MIC50 HemolysisCompound (uM) (at 100 uM) AA 7.5 <10% AB 7.5 <10% A1 7.5 <10% A3 7.5<10% A4 7.5 <10% A7 2.5 <10% B4 2.5 <10% B7 7.5 <10% 7 7.5 50% 8 17.5<10% 9 17.5 <10%

The antimicrobial test results for the Library 5 compounds showed someinteresting trends and various degrees of activities against B. subtilis(Table 4). In general, increasing in bulkiness (ring size) on one sidechain of triazine compounds increased the antimicrobial activity, wheresize 7 and 8 seemed to be optimal. If the ring size was increased to 12(A5), activity decreased almost 8-fold comparing to size 7 or 8compounds. Interestingly, triazine compounds with two amantadine groupshad much less antimicrobial activity compared with compounds having onlyone amantadine group. On the other hand, increasing the amine chainlength did not show obvious trend, with some slight increase in activityamong some compounds. In addition, quaternary ammonium cation also didnot have any large effect on improving the activity.

TABLE 4 MIC50 results from library 5.

Compound MIC50 (uM) A1 16.4 A2 7.9 A3 4.6 A4 2.0 A5 15.9 A6 7.6 A7 5.5A8 39 A9 4.4 A10 5 C1 10.6 C2 2.7 C3 3.7 C4 3.5 a9 5.3 b9 7 c9 5.6 d95.7 e9 5.9 f9 4.7 g9 5.9 h9 N/A C5-S 22.5

Finally, three compounds, A7 from Library 3, A4 and C2 from Library 5,were selected and their antimicrobial activity and hemolytic activitywere tested against different bacteria, and sheep blood cells (Table 5).The results showed that these compounds were generally more selectiveagainst gram-positive bacteria, and were not highly hemolytic even athigh concentration.

TABLE 5 MIC50 and HC50 of compounds A4 (from library 5), C2 (fromlibrary 5) and A7 (from library 3) against different bacteria and sheepblood cells. MIC50 (uM) MIC50 (uM) MIC50 (uM) A4 C2 A7 A. baumannii 8422 53 B. anthracis 86 21 41 E. coli D31 546 200 618 E. faecalis 320 142266 S. aureus 70 17 68 HC50 A4 HC50 C2 HC50 A7 Hemolysis >>2 mg/ml >>2mg/ml >>2 mg/ml

Example 3

Calcein Dye Leakage

To test whether the active triazine compounds kill bacteria viadisrupting membrane integrity, a calcein dye leakage experiment wascarried out with compound C2 with ampicillin and one of the triazinecompounds with little antimicrobial activity (h-9) as control. Themethod of preparation of dye-encapsulated vesicles has been reported indetail. Lasch, V.; Weissig, M. Brandl in Liposomes: a Practical ApproachEds.: Taylor, K. M. G.; Craig, D.Q.M. Oxford University Press, Oxford,2003; pp. 10-12. The results are graphically presented in FIG. 1, whichpresents the results of calcein leakage induced by 1% Triton, C2 (50μM), h9 (90 μM) and ampicillin (70 μM). The fraction of leakage wascalculated from the fluorescence intensity at 515 nm, with 100% leakagecalibrated by addition of 1% Triton X-100.

From the results, C2 was found to cause 100% dye leakage at 50 μM. Theresults confirmed that the antimicrobial active triazine compound couldcause dye leakage from negatively charged1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG)lipid vesicles, while neither ampicillin nor compound C2 caused dyeleakage.

REFERENCES

-   1. Zasloff, M. 2002. Antimicrobial peptides of multicellular    organisms. Nature 415(6870):389-95.-   2. Boman, H. G. 2003. Antibacterial peptides: basic facts and    emerging concepts. J. Intern. Med. 254(3):197-215.-   3. Medzhitov, R. and C. Janeway, Jr. 2000. Innate immune    recognition: mechanisms and pathways. Immunol. Rev. 173:89-97.-   4. Brogden, K. A., Ackermann, M., McCray, P. B., and    Tack, B. F. 2003. Antimicrobial peptides in animals and their role    in host defences. Int J Antimicrob Agents 22(5):465-78.-   5. Hancock, R. E. 1999. Host defence (cationic) peptides: what is    their future clinical potential? Drugs 57(4):469-73.-   6. Fischetti, V. A. 2003. Novel method to control pathogenic    bacteria on human mucous membranes. Ann. N.Y. Acad. Sci. 987:207-14.-   7. Yeaman, M. R. and Yount, N.Y. 2003. Mechanisms of antimicrobial    peptide action and resistance. Pharmacol Rev. 55:27-55.-   8. Str{acute over (ç)}m, M. B., Haug, B. E., Skar, M. L., Stensen,    W., Stiberg, T. and Svendsen, J. S. 2003. The pharmocophore of short    cationic antibacterial peptides. J. Med. Chem. 46(9):1567-1570.-   9. Liu, Z., Brady, A., Young, A., Rasimick, B., Chen, K., Zhou, C.    and Kallenbach, N. R. 2007. Length effects in antimicrobial peptides    of the (RW)n Series. Antimicrobial Agents and Chemotherapy 51    (2):597-603.-   10. Arvidsson, P. I., Frackenpohl, J., Ryder, N. S., Leichty, B.,    Petersen, F., Zimmermann, H., Camenisch, G. P., Woessner, R. and    Seebach, D. 2001. On the antimicrobial and hemolytic activities of    amphiphilic β-peptides. ChemBioChem 2(10):771-773.-   11. Goodson, B., Ehrhardt, A., Ng, S., Nuss, J., Johnson, K.,    Giedlin, M., Yamamoto, R., Moos, W. H., Krebber, A., Ladner, M.,    Giacona, M. B., Vitt, C. and Winter, J. 1999. Characterization of    novel antimicrobial peptoids. Antimicrobial Agents and Chemotherapy    43(6):1429-1434.-   12. Rennie, J., Amt, L., Tang, H., Nuesslein, K. and    Tew, G. N. 2005. Simple oligomers as antimicrobial peptide    mimics. J. Industrial Microbiology & Biotech. 32(7):296-300.-   13. Silen, J. L., Lu, A. T., Solas, D. W., Gore, M. A., Maclean, D.,    Shah, N. H., Coffin, J. M., Bhinderwala, N. S., Wang, Y.,    Tsutsui, K. T., Look, G. C., Campbell, D. A., Hale, R. L., Navre, M.    and DeLuca-Flaherty, C. R. 1998. Screening for novel antimicrobials    from encoded combinatorial libraries by using a two-dimensional agar    format. Antimicrobial Agents and Chemotherapy 42(6): 1447-1453.-   14. Giacometti, A., Cirioni, O., Greganti, G., Quarta, M. and    Scalise, G. 1998. In vitro activities of membrane-active peptides    against gram-positive and gram-negative aerobic bacteria.    Antimicrobial Agents and Chemotherapy 42(12):3320-3324.-   15. Giacometti, A., Cirioni, O., Barchiesi, F., Del Prete, M. S, and    Scalise, G. 1999. Antimicrobial activity of polycationic peptides.    Peptides 20(11):1265-1273.-   16. Huang, H. W. 2000. Action of antimicrobial peptides: two-state    model. Biochemistry 39(29):8347-8352.

From the foregoing description, various modifications and changes in thecompositions and methods of this invention will occur to those skilledin the art. All such modifications coming within the scope of theappended claims are intended to be included therein.

It is further understood that all base sizes or amino acid sizes, andall molecular weight or molecular mass values, given for nucleic acidsor polys-triazine compounds are approximate, and are provided fordescription.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

1. A s-triazine compound having a formula II:

wherein each of R^(2f), and R^(2g) is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is selected from 3-13membered cycloalkyl ring; or a pharmaceutically acceptable salt thereof;and stereoisomers, isotopic variants and tautomers thereof.
 2. Acompound of claim 1 wherein R³ is cyclopropyl or cyclobutyl.
 3. Acompound of claim 1 wherein R³ is cycloheptyl.
 4. A s-triazine compoundhaving a formula II:

wherein each of R^(2f), and R^(2g) is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is R⁴; and R⁴ is selectedfrom substituted or unsubstituted

or a pharmaceutically acceptable salt thereof; and stereoisomers,isotopic variants and tautomers thereof.
 5. A s-triazine compound havinga formula II:

wherein each of R^(2f), and R^(2g) g is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is C₁-C₆ alkyl,unsubsituted or substituted with acyl, acylamino, acyloxy, alkoxy,alkoxycarbonyl, alkoxycarbonylamino, amino, azido, cyano, halogen,hydroxyl, heteroaryl, keto, nitro, thioalkoxy, alkyl-S(O)—, aryl-S(O)—,alkyl-S(O)₂—, or aryl-S(O)₂—; or a pharmaceutically acceptable saltthereof; and stereoisomers, isotopic variants and tautomers thereof. 6.A s-triazine compound having a formula II:

wherein each of R^(2f), and R^(2g) is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is unsubstituted phenyl;or a pharmaceutically acceptable salt thereof; and stereoisomers,isotopic variants and tautomers thereof.
 7. A compound selected from:

or a pharmaceutically acceptable salt thereof; and stereoisomers,isotopic variants and tautomers thereof; and provided the compound isnot h9.
 8. A compound of claim 7 wherein the compound is

or a pharmaceutically acceptable salt thereof; and stereoisomers,isotopic variants and tautomers thereof.
 9. A compound of any one ofclaim 1, 4, 5, 6, or 7 wherein the compound is a quaternary salt.
 10. Apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a pharmaceutically effective amount of a compound of any oneof claim 1, 4, 5, 6, or
 7. 11. The pharmaceutical composition of claim10, wherein the carrier is selected from a topical carrier, an oralcarrier, or a parenteral carrier.
 12. A compound according to claim 5,wherein R³ is substituted or unsubstituted methyl, ethyl, n-propyl, orn-butyl.
 13. A s-triazine compound having a formula II:

wherein each of R^(2f), and R^(2g) is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is phenyl, optionallysubstituted with one or more substituents independently selected fromhalo, alkoxy, or alkyl; or a pharmaceutically acceptable salt thereof;and stereoisomers, isotopic variants and tautomers thereof.
 14. Acompound according to claim 13, wherein R³ is phenyl, substituted withCl, OMe, methyl, ethyl, n-propyl, or n-butyl.
 15. A s-triazine compoundhaving a formula II:

wherein each of R^(et), and R^(2g) is independently selected fromhydrogen, and substituted or unsubstituted C₁-C₆ alkyl; or R^(2f) andR^(2g) may join together to form a cycloheteroalkyl ring of 3-7 atoms; mis 0, 1, 2, or 3; and n is 2, 3, or 4; and R³ is heteroaryl, optionallysubstituted with one or more substituents independently selected fromhalo, alkoxy, or alkyl; or a pharmaceutically acceptable salt thereof;and stereoisomers, isotopic variants and tautomers thereof.
 16. Acompound according to claim 15, wherein R³ is pyridyl, unsubstituted orsubstituted with Cl, OMe, methyl, ethyl, n-propyl, or n-butyl.
 17. Amethod for treating, ameliorating or managing a disease or conditionwhich comprises administering to a patient in need of such treatment,amelioration or management, a therapeutically effective amount of apharmaceutical composition of claim 10, wherein the disease or conditionis or results from a bacterial infection.